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Plant and Cell Physiology, 2001, Vol. 42, No. 6 635-641
© 2001 Oxford University Press

A TGACGT Motif in the 5'-Upstream Region of {alpha}-Amylase Gene from Vigna mungo is a cis-Element for Expression in Cotyledons of Germinated Seeds

Daisuke Yamauchi1 Department of Biological Sciences, Tokyo Metropolitan University, Minami-ohsawa 1-1, Hachioji, Tokyo, 192-0397 Japan

{alpha}-Amylase is expressed at high levels in cotyledons of germinated seeds of Vigna mungo. The mRNA for {alpha}-amylase appeared in cotyledons of the seeds at 1 d after imbibition started (DAI). Two TGACGT motifs at –445 and at –125 in the promoter region of the gene interacted with nuclear proteins from cotyledons of dry seeds and the activities were detected until 3 DAI. A transient assay with particle bombardment showed that the downstream region from –135 in the promoter was required for high level expression in the cotyledons and the activity was reduced by mutation of the TGACGT motif at –125. The activities to bind the TGACGT motifs were detected in the axes of the seeds at 1 DAI but disappeared at 4 DAI, although the mRNA for {alpha}-amylase in the axes appeared at 4 DAI and increased in level by 6 DAI. A transient assay experiment showed that a positive regulatory element for the expression in the axes was located in the region from –630 to –453. These results indicated that the TGACGT motif at –125 was required for high level expression of the gene in the cotyledons of the germinated seeds.

1 Corresponding author: E-mail: yamauchi-daisuke@c.metro-u.ac.jp; Fax: +81-426-77-2559.


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